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Promoting extracellular matrix remodeling via ascorbic acid enhances the survival of primary ovarian follicles encapsulated in alginate hydrogels

Authors

  • David Tagler,

    1. Department of Chemical and Biological Engineering, McCormick School of Engineering, Northwestern University, Evanston, Illinois
    2. Institute for BioNanotechnology in Medicine (IBNAM), Northwestern University, Chicago, Illinois
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  • Yogeshwar Makanji,

    1. Feinberg School of Medicine, Northwestern University, Chicago, Illinois
    2. Center for Reproductive Science (CRS), Northwestern University, Evanston, Illinois
    3. Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, Illinois
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  • Tao Tu,

    1. Department of Chemical and Biological Engineering, McCormick School of Engineering, Northwestern University, Evanston, Illinois
    2. Institute for BioNanotechnology in Medicine (IBNAM), Northwestern University, Chicago, Illinois
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  • Beatriz Peñalver Bernabé,

    1. Department of Chemical and Biological Engineering, McCormick School of Engineering, Northwestern University, Evanston, Illinois
    2. Institute for BioNanotechnology in Medicine (IBNAM), Northwestern University, Chicago, Illinois
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  • Raymond Lee,

    1. Feinberg School of Medicine, Northwestern University, Chicago, Illinois
    2. Center for Reproductive Science (CRS), Northwestern University, Evanston, Illinois
    3. Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, Illinois
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  • Jie Zhu,

    1. Feinberg School of Medicine, Northwestern University, Chicago, Illinois
    2. Center for Reproductive Science (CRS), Northwestern University, Evanston, Illinois
    3. Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, Illinois
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  • Ekaterina Kniazeva,

    1. Department of Chemical and Biological Engineering, McCormick School of Engineering, Northwestern University, Evanston, Illinois
    2. Institute for BioNanotechnology in Medicine (IBNAM), Northwestern University, Chicago, Illinois
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  • Jessica E. Hornick,

    1. Feinberg School of Medicine, Northwestern University, Chicago, Illinois
    2. Center for Reproductive Science (CRS), Northwestern University, Evanston, Illinois
    3. Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, Illinois
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  • Teresa K. Woodruff,

    1. Feinberg School of Medicine, Northwestern University, Chicago, Illinois
    2. Center for Reproductive Science (CRS), Northwestern University, Evanston, Illinois
    3. Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, Illinois
    4. Chemistry of Life Processes Institute (CLP), Northwestern University, Evanston, Illinois
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  • Lonnie D. Shea

    Corresponding author
    1. Department of Chemical and Biological Engineering, McCormick School of Engineering, Northwestern University, Evanston, Illinois
    2. Institute for BioNanotechnology in Medicine (IBNAM), Northwestern University, Chicago, Illinois
    3. Center for Reproductive Science (CRS), Northwestern University, Evanston, Illinois
    4. Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Chicago, Illinois
    5. Chemistry of Life Processes Institute (CLP), Northwestern University, Evanston, Illinois
    • Correspondence to: L.D. Shea.

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  • Authors contributed equally to manuscript.

ABSTRACT

The in vitro growth of ovarian follicles is an emerging technology for fertility preservation. Various strategies support the culture of secondary and multilayer follicles from various species including mice, non-human primate, and human; however, the culture of early stage (primary and primordial) follicles, which are more abundant in the ovary and survive cryopreservation, has been limited. Hydrogel-encapsulating follicle culture systems that employed feeder cells, such as mouse embryonic fibroblasts (MEFs), stimulated the growth of primary follicles (70–80 µm); yet, survival was low and smaller follicles (<70 µm) rapidly lost structure and degenerated. These morphologic changes were associated with a breakdown of the follicular basement membrane; hence, this study investigated ascorbic acid based on its role in extracellular matrix (ECM) deposition/remodeling for other applications. The selection of ascorbic acid was further supported by a microarray analysis that suggested a decrease in mRNA levels of enzymes within the ascorbate pathway between primordial, primary, and secondary follicles. The supplementation of ascorbic acid (50 µg/mL) significantly enhanced the survival of primary follicles (<80 µm) cultured in alginate hydrogels, which coincided with improved structural integrity. Follicles developed antral cavities and increased to diameters exceeding 250 µm. Consistent with improved structural integrity, the gene/protein expression of ECM and cell adhesion molecules was significantly changed. This research supports the notion that modifying the culture environment (medium components) can substantially enhance the survival and growth of early stage follicles. Biotechnol. Bioeng. 2014;111: 1417–1429. © 2013 Wiley Periodicals, Inc.

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